Description:

Tumour Necrosis Factor alpha (TNFa), is an inflammatory cytokine produced by macrophages/monocytes during acute inflammation and is responsible for a diverse range of signaling events within cells, leading to necrosis or apoptosis. The protein is also important for angiogenesis that is critical to the growth, progression, and metastasis of solid tumors. Furthermore, TNFa is associated with obesity. It is chronically elevated in adipose tissues of obese rodents and humans and may represent an important link between obesity and insulin resistance. In both obese mice and humans, TNFa is overexpressed in adipose tissue. TNFa inhibits insulin signaling, at least in part by blocking insulin receptor tyrosine kinase activity and inducing serine phosphorylation of insulin receptor substrate-1 (IRS-1). However, it is unclear what the physiological stimulator of TNFa production by adipocyte during obesity is and how IRS-1 inhibits the tyrosine kinase activity of the insulin receptor after TNFa treatment of the cells. A better understanding of the connection(s) between the TNFa and the insulin signaling pathways could be important to find a cure for the state of insulin resistance observed during obesity.

Principle:

The ELISA is based on the principle of a solid phase enzyme-linked immunosorbent assay. The assay utilizes a anti-rat cytokine antibody for immobilization on the microtiter wells and anti-rat cytokine antibodies along with streptavidin conjugated to horseradish peroxidase (HRP) for detection. The test sample is allowed to react simultaneously with the two antibodies, resulting in the molecules of the cytokine being sandwiched between the solid phase and enzyme-linked antibodies. After incubation, the wells are washed to remove unbound-labeled antibodies. A HRP substrate, TMB, is added to result in the development of a blue color. The color development is then stopped with the addition of Stop Solution changing the color to yellow. The concentration of the cytokine is directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450 nm.

 

 

 

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Signosis 是一家专业的生物分析供应商，致力于基于微孔板分析技术的生命科学研究产品的开发、生产和销售。Signosis 成立于2007年，坐落于世界研发中心--美国硅谷的中心地带。公司专注于细胞因子、转录因子、miRNA及疾病标志物等重要调控因子和标志物的研究，基于自有专利技术，开发完成400多种先进而独特的用于发现、筛选和分析的生命科学研究产品。Signosis 产品销售遍布北美、欧洲、亚洲及中东等主要国家和地区，客户涵盖美国国立卫生研究院、哈佛大学、耶鲁大学等著名研究机构以及众多制药公司